Abstract:
:RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function.
journal_name
Proc Natl Acad Sci U S Aauthors
Kittler R,Pelletier L,Ma C,Poser I,Fischer S,Hyman AA,Buchholz Fdoi
10.1073/pnas.0409861102keywords:
subject
Has Abstractpub_date
2005-02-15 00:00:00pages
2396-401issue
7eissn
0027-8424issn
1091-6490pii
0409861102journal_volume
102pub_type
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